The lateral resolution is about 50 μm. This technique paves the way for applying dual-comb in remote sensing and imaging.We propose and experimentally demonstrate single-pixel photon counting imaging based on dual-comb interferometry at 1550 nm. Different from traditional dual-comb imaging, this approach enables imaging at the photon-counting regime by using single-photon detectors combined with a time-correlated single-photon counter to record the returning photons. The illumination power is as low as 14 pW, corresponding to 2.2 × 10 The development of single-photon detectors has revolutionized photon-counting laser ranging and imaging in speed and sensitivity ,7,8,9,10−3 photons/pulse and lateral resolution of about 50 μm. Compared to a conventional dual-comb imaging system that uses a normal fast photodetector with the sensitivity at about milliwatts to microwatts [In this paper, we employed the dual-comb Fourier transform spectroscopy to retrieve the single-photon level signal, which carries spectrally encoded spatial information around 1550 nm. The experiment was conducted at an extremely low photon flux level, and we utilized a single-pixel single-photon detector together with a time-correlated single-photon counter (TCSPC) to detect the temporal interferogram waveform of the target-encoded reflected photons. A two-dimensional image was obtained by scanning the target with the dual optical frequency comb at a photon flux of 2.2 × 10crowatts ,13,15,16frep1 (=25 GHz) and frep2 , respectively. The comb carrier-envelope frequency was centered at feo = 192.8 THz, and one of the combs was shifted by Δfeo = 40 MHz using an acousto-optic modulator for avoiding frequency aliasing in dual-comb measurements [N = 120 comb lines. Thus, the frequency of the nth comb line of a comb could be written as feo ± n·frep (or feo + Δfeo ± n·frep), where n ≤ N/2 (n is an integer). Then, the two combs were combined by a 50:50 fiber coupler and collimated to the free-space by a fiber collimator after passing through a variable fiber attenuator and a fiber circulator. For spectral encoding imaging, the dual-comb beam was spatially dispersed with a pair of gratings (600 lines/mm) and then focused on a target in vertical direction by a cylindric lens with a focal length of 100 mm. The target was a commercialized resolution test chart with positive patterns  fabricated by vacuum depositing chrome patterns on the glass substrate as a reflective film. The target was placed on the focal plane of the cylinder lens. Consequently, a long strip-shaped area on the target was illuminated. The spatially dispersed light established a one-to-one correspondence between the spatial locations and the spectral elements  in a one-dimensional spectrum. Therefore, mechanically scanning the target in the direction vertical to the dispersed incident plane led to a two-dimensional (2D) image of a portion of the target . In our case, the AWG output  was set to Δfrep/8 (=62.5 kHz), and correspondingly, the time window was eight times the dual-comb period.Dual-comb spectroscopy due to its unique spectral properties such as broadband spectra, uniformly spaced frequency distribution, long-term coherence, high resolution, and stability, has been demonstrated to be a powerful tool for many different areas. The time-correlated single-photon counting technique, combined with a single-photon detector, can improve the dual-comb imaging system photon sensitivity and can reduce counting noise, and enhance the image quality. Based on this idea, we designed the photon-counting dual-comb interferogram imaging experiment. The experimental setup is sketched in urements . The twoWe first used an intensedual-comb beam (with weak attenuation) for testing and measured spectrum-encoded images with an optical spectrum analyzer . The wavelength resolution of the spectrometer was set to 0.05 nm, and the sampling resolution was set to 0.01 nm. The laser output power at the fiber collimator was about 1.745 mW and the returning laser power acquired by the spectrometer varied from 6 to 46 μW when scanning the different locations on the target. We scanned the target vertically with a step size of 0.05 mm close to the vertical beam size at the focus spot. For example, With the spectrum-encoding technique, we could get 2D spatial information of the targeted area. An example is shown in −3 photons/pulse. At this point, the returning photon flux was too weak for the optical spectrum analyzer to be detected. We replaced the optical spectrum analyzer with a single-photon detector for the 2D dual-comb imaging at photon-counting level. Firstly, we used a superconducting nanowire single-photon detector  in the system as the photon receiver. The photon detection probability of the SNSPD was about 78% around 1550 nm, and the dark count noise was about 32 counts/s. The counting rate of the detector varied from 8.3 × 104 counts/s to 8 × 105 counts/s when scanning the light across the target. The output signal from the single-photon detector was sent to the stop channel of the TCSPC. repf/8, we could observe 8 periods within 16 μs in the photon-counting dual-comb interferograms as shown in tFFT frep1/Δfrep, where tFFT was the apodization window of the interferometric data and 1.78 was a factor for triangular apodization [−18 W). Consequently, the system will be a favorable platform for remote detection where the returning photon flux is quite low, such as in astronomical observation [Then, the output laser power at the fiber collimator was attenuated to a few photons/pulse level. The light power after the collimator was 14 pW, corresponding to 2.2 × 10dization ,22. In tervation .We implemented the dual-comb imaging at photon-counting level by scanning the target vertically in a stepwise manner. A 2D dual-comb spectrum-encoding image with spatial resolution of ≈50 μm is shown in 3 counts/s. The 2D dual-comb image acquired by the InGaAs-APD detector is shown in Furthermore, we compared dual-comb photon-counting imaging with single-photon detectors operating in different modes. We replaced the SNSPD with an InGaAs-APD . Different from the SNSPD, which was operated in continuous mode, the InGaAs-APD was operated in gated Geiger-mode to reduce the dark count noise and the afterpulsing noise of the detector itself. The photon detection probability of the InGaAs-APD was about 7%. The repetition rate of the gating pulse was 1.25 GHz, and the dark count noise was about 1.2 × 10In conclusion, by combining a single-pixel single-photon detector together with a time-correlated single-photon counter (TCSPC) in a dual-comb target-encoded imaging system, we achieved two-dimensional imaging at the single-photon level with both high precision and high detection sensitivity at 1550 nm. The 2D image was recovered by line scanning the target with the dual-comb laser light at the few photon level, showing great potential in the remote imaging where the transmission loss could not be ignored. Unlike in general spectrum encoding, there is no dispersing element in the detection part in the dual-comb Fourier transform spectroscopy imaging system, enhancing the mechanic stability of the imaging system. This method may have the potential to find widespread applications in nanomaterial imaging where the incident light should be very weak to avoid the damage of the sample.


Critical illness in the intensive care unit (ICU) has been a global health priority. Systemic nutritional status has turned out to be related to the prognosis of critically ill patients. The albumin-globulin ratio (AGR) has been reported to be a novel prognostic factor of many diseases. This study is aimed at investigating whether the AGR could predict the mortality risk in critically ill patients.  We enrolled 582 adult patients admitted to the respiratory intensive care unit (RICU). We collected the clinical and laboratory data. X-tile software was used to determine the optimal cut-off values for the AGR. Patients were divided into three groups according to the AGR . Kaplan-Meier analysis was used for survival analysis. A Cox proportional hazard model was applied to the univariate and multivariate analyses for the potential predictors associated with survival. p = 0.02); HR: 0.332 (0.166-0.665) (p = 0.002)).  Our present study showed that the AGR was related to the 28-day survival of critically ill patients in the RICU. The rate of pneumonia in the low AGR group was significantly higher than that in the other groups. Patients with a lower AGR present an increased risk of 28-day mortality compared to patients with a higher AGR. Cox regression analysis showed that the AGR might be an independent predictor of prognosis to 28-day survival in critically ill patients in the RICU. Medium and high AGR values remained independently associated with better 28-day survival than low AGR values (HR: 0.484 (0.263-0.892) ( The AGR might be an independent predictor of prognosis in critically ill patients. Critical illnesses in the intensive care unit (ICU), such as sepsis, could be a global health priority , 2. BeinSerum albumin is commonly used as a surrogate of nutritional status, and serum globulin is cited to assess the severity of chronic inflammation . HypoalbWe conducted a retrospective cohort study of 582 adult patients admitted to the respiratory intensive care unit (RICU) in the College of Pulmonary and Critical Care Medicine, Chinese PLA General Hospital, from 1 July 2008 to 31 December 2017. We collected the clinical data of these patients for retrospective analysis. The inclusion criteria of the study were as follows: (1) adult patients (age ≥ 18 years old), (2) patients with a laboratory examination of serum albumin and serum total protein, (3) patients with a laboratory examination of liver function, and (4) patients with a RICU stay of 2 to 100 days. The exclusion criteria of the study were as follows: (1) patients younger than 18 years old and (2) patients with insufficient AGR data. The primary outcome of the study was 28-day mortality.The baseline data of the patients included were collected during the stay in the RICU. The AGR was calculated by the following formula: AGR = serum albumin/ .X-tile 3.6.1 software  was used to determine the optimal cut-off values for the AGR. IBM SPSS Statistics 21.0 software  was applied to complete the statistical analysis. Data are described as numbers (percentages) for categorical variables, means ± standard deviations for normally distributed variables, and medians (interquartile ranges) for skewed distributed variables. Patients were divided into three groups according to the AGR. Χ2 tests were used to analyze the relationship between the clinical parameters and AGR. Kaplan-Meier analysis was used for survival analysis. A Cox proportional hazard model was applied to the univariate and multivariate analyses for the potential predictors associated with survival. The results were considered statistically significant with a two-sided p value <0.05.μmol/L, and 6.8 μmol/L, respectively. Among these 582 patients, 267 (45.9%) patients had a comorbidity of pneumonia, and 93 (15.8%) patients had a comorbidity of respiratory failure. The average length of stay (LOS) in the RICU was 16 days. The 28-day mortality for the entire cohort was 10.0% . The AGR of these patients ranged from 0.2 to 2.24, with a median value of 1.06. Data on liver function-related factors, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), and direct bilirubin (DBIL), were collected. The average ALT, AST, TBIL, and DBIL levels were 34 U/L, 37 U/L, 14.8 as 10.0% .X-tile software to determine the optimal cut-off value for the AGR of 28-day mortality. X-tile is a robust graphical tool verified by Yale University [X-tile could be convincing. The optimal cut-off values determined by X-tile for the AGR were 0.8 and 1.1 (p < 0.001) (p = 0.001).We used iversity , and the and 1.1 . Accordi< 0.001) . The ratp = 0.004. Kaplan-Meier analysis showed that the low AGR group had the worst 28-day survival, and the high AGR group had the best 28-day survival, indicating that the AGR might be related to the prognosis of critically ill patients in the RICU.p = 0.036) and 0.330 (0.166-0.655) (p = 0.002), respectively. Meanwhile, elevated DBIL and TBIL and age ≥ 65 years old indicated a correlation with worse 28-day survival (HR: 2.232 (1.306-3.815) (p = 0.003), HR: 2.016 (1.156-3.517) (p = 0.014), and HR: 2.19 (1.231-3.896) (p = 0.008)), respectively). In the multivariate analysis, medium and high AGR values remained independently associated with better 28-day survival (HR: 0.484 (0.263-0.892) (p = 0.02) and HR: 0.332 (0.166-0.665) (p = 0.002)) than low AGR values. Age exceeding 65 years remained an independent factor for worse 28-day survival (HR: 2.196 (1.23-3.92) (p = 0.009))  of 0.523 (0.285-0.959) ( 0.009)) .To the best of our knowledge, this is the first study to examine the prognostic value of the AGR in terms of 28-day survival in critically ill patients. Our present study showed that the AGR was related to the 28-day survival of critically ill patients in the RICU. The rate of pneumonia in the low AGR group was significantly higher than that in the other groups. Patients with a lower AGR present an increased risk of 28-day mortality compared to patients with a higher AGR. Cox regression analysis showed that the AGR might be an independent predictor of prognosis to 28-day survival in critically ill patients in the RICU.The condition of patients in the ICU is usually poor, causing a serious economic burden to society and family. Therefore, it is very important to find novel markers in the prognosis of critical illness, which might be helpful in targeted therapy. Agnello et al. showed that monocyte distribution width was associated with sepsis in the ICU . In addiα (TNF-α) and interleukin-6 (IL-6), turned out to be responsible for the altered albumin in critically ill patients [The nutritional status of critically ill patients will be affected due to long-term bed stays and inflammation. Therefore, nutritional status was associated with the prognosis of critically ill patients. Nutritional risk has been proven to be a predictor of 28-day and ICU mortality . Many stpatients . The etipatients .α [The AGR has drawn increasing attention for its potential role as a predictor in many diseases. The AGR was assumed to be correlated with prognosis in colorectal cancer , lung caα . Undurtiα . In our Our study showed that the AGR might be an independent predictor of the prognosis to 28-day survival in critically ill patients. A lower AGR indicated a worse prognosis in critically ill patients. We might find an efficient way to increase albumin and the AGR; so, we could give critically ill patients an encouraging prognosis.There are some limitations to our retrospective study. First, this is a single-centre retrospective study, and the sample size is relatively small compared to that in a multicentre study. In addition, there might be selection bias, which is the defect in most single-centre retrospective studies. Second, we did not collect the AGR during the whole stay in the RICU, and we cannot accurately analyze the trend of the AGR with disease progression. Finally, our study was a clinical study and did not examine the exact mechanism by which the AGR affects the prognosis of critically ill patients. More studies of the mechanisms of the AGR and disease are needed in the future.In the present study, we showed that the AGR was correlated with the 28-day survival of critically ill patients in the RICU. Patients with a lower AGR present an increased risk of 28-day mortality compared to patients with a higher AGR. AGR might be an independent predictor of prognosis to 28-day survival in critically ill patients. Multi-institutional studies are needed to confirm our present findings. How to adjust the therapeutic strategies of critically ill patients with a poor prognosis determined by the AGR might be a subject worth studying in the future.


Here, we report an alginate-confined peroxygenase-CLEA to catalyse the enantioselective epoxidation of  cis-β-methylstyrene under non-aqueous reaction conditions. Product titres of up to 48 mM and excellent enzyme turnovers of 96 000 have been achieved.An alginate-confined peroxygenase-CLEA has been prepared for the stereoselective epoxidation of  P450 monooxygenases reductively activate molecular oxygen to form the catalytically active oxyferryl-heme species. Peroxygenases form this species directly from partially reduced oxygen species (peroxides) and thereby circumvent the complex molecular architectures of P450 monooxygenases  concentration is of utmost importance to increase the economic viability and environmental friendliness of such biocatalytic reactions.5 in which an aqueous, biocatalyst-containing layer is contacted with a hydrophobic, organic layer serving as substrate reservoir and product sink. To alleviate possible phase transfer rate limitations of this system, intensive mechanical stirring is needed, which however, also may impair the stability of the biocatalyst.6One common approach to increase the substrate loading is to use the so-called two liquid phase approach7–12 In previous works, we could demonstrate that immobilised peroxygenases in principle can even be applied in neat  reaction media.9 A drawback of this approach, however, was the very poor specific activity of the immobilised enzyme, possibly due to a combination of activity losses of the enzyme during immobilisation and further activity losses originating from dehydratation of the enzyme surface.The latter issue can be addressed by immobilising the biocatalyst to a heterogeneous carrier material and thereby physically protecting the enzyme. While a limited number of studies report immobilisation of peroxygenases, this technique is not fully explored yet for this enzyme class.8,11,13Encapsulating enzymes in alginate matrices may represent an elegant compromise. The so-confined enzymes are mechanically stabilised while still situated in a micro-aqueous environment.Agrocybe aegerita (rAaeUPO)14–16 as catalyst for the epoxidation of styrene and cis-β-methylstyrene  and 11.4% of the peroxidase activity, as judged by the ABTS oxidation activity, was found back in the immobilisates. It is worth mentioning here that rAaeUPO immobilisation also increased its storage stability. While the free enzyme completely lost its catalytic activity after 12 days storage at room temperature, the immobilised version exhibited at least 80% of its initial activity even after two weeks  while with the free enzyme it was only 3 mM h−1. A plausible explanation for this may be the higher surface area of the reactions using alginate-immobilised rAaeUPO, largely eliminating the diffusion rate limitation of tBuOOH and/or cis-β-methylstyrene into the aqueous reaction phase . With the immobilised enzyme the product accumulation rate was approx. 10 mM hR,3S)-2-methyl-3-phenyloxirane has been synthesised within 4 h corresponding to a turnover number (TN = molProduct × molAaeUPOr−1) for the enzyme of 60 000 and an average (over 4 h) turnover frequency of 4.1 s−1. It should be mentioned here, that under these reaction conditions several side products such as benzaldehyde and phenylacetone were observed (vide infra).Overall, in this experiment, approx. 30 mM of enantiomerically pure  of enantiomerically pure -2-methyl-3-phenyloxirane have been synthesised within 72 h. This corresponds to excellent TN of 96 000 for the biocatalyst. This catalytic performance also favourably compares to other enzyme systems such as flavin-dependent monooxygenases.24–27 Admittedly, the current substrate scope of the proposed immobilised rAaeUPO preparation is rather limited, preliminary experiments on styrene epoxidation indicate a similar catalytic potential for this substrate -2-methyl-3-phenyloxirane. As shown in via distillation. The second-largest contributor (24%) is the enzyme preparation. The latter is mostly comprised of the alginate beads and buffer .To obtain a first overview over the environmental impact of the reaction system established in this study, we used Sheldon's E-factorAaeUPO loading in the alginate beads. For example using alginate-in-oil emulsions for the bead preparation will certainly improve the rAaeUPO loading.30 Also the E-factor contribution of the oxidant and its by-product tBuOH, respectively, can be reduced significantly when co-immobilising the formate oxidase from Aspegillus oryzae (AoFOX) to use methanol as sacrificial electron donor for the in situ generation of H2O2.31–34 Implementing this system will eliminate the tBuOH contribution to the E-factor (8.5 kg kg−1) and reduce it to approx. 0.11 kgCO2 kgProduct−1.We are convinced that further optimisation of the immobilisation protocol will improve the rcis-β-methylstyrene to synthesise enantiomerically pure epoxides. In terms of catalyst efficiency , the current system outperforms comparable reaction systems using chemical catalysts,35 P450 monooxygenases36 or other established enzymatic systems,26,27,37 also compared to our previous efforts using immobilised rAaeUPO.9 We are convinced that further optimisation will bring this approach to maturity and will establish an economically and environmentally feasible reaction system.Overall, in this contribution we have established alginate-confined peroxygenase-CLEAs as practical enzyme preparations for the selective epoxidation of styrene derivates such as Financial support by the China Scholarship Council, the from the German Academic Exchange Service and the German Academic Scholarship Foundation as well as the European Research Council (ERC Consolidator Grant No. 648026) is gratefully acknowledged.F. E. H. N., Y. W. and M. P. have performed the experiments and analysed the results. The study was conceptualised by M. H., M. A. and F. H. All authors contributed to the manuscript writing.There are no conflicts to declare.CC-057-D1CC01868J-s001


In two-sample Mendelian randomization (MR) studies, sex instrumental heterogeneity is an important problem needed to address carefully, which however is often overlooked and may lead to misleading causal inference.Q test to examine sex genetic similarity and heterogeneity in instrumental variables (IVs) of exposures. Simulation was further performed to explore the influence of sex instrumental heterogeneity on causal effect estimation in sex-specific two-sample MR analyses. Furthermore, we chose breast/prostate cancer as outcome and four anthropometric traits as exposures as an illustrative example to illustrate the importance of taking sex heterogeneity of instruments into account in MR studies.We first employed cross-trait linkage disequilibrium score regression (LDSC), Pearson’s correlation analysis, and the Cochran’s Q test also displayed sex heterogeneity for some instruments. When applying sex-specific instruments, significant discrepancies in the magnitude of estimated causal effects were detected for body mass index (BMI) on breast cancer (P = 1.63E-6), for hip circumference (HIP) on breast cancer (P = 1.25E-20), and for waist circumference (WC) on prostate cancer (P = 0.007) compared with those generated with sex-combined instruments.The simulation definitively demonstrated that sex-combined IVs can lead to biased causal effect estimates in sex-specific two-sample MR studies. In our real applications, both LDSC and Pearson’s correlation analyses showed high genetic correlation between sex-combined and sex-specific IVs of the four anthropometric traits, while nearly all the correlation coefficients were larger than zero but less than one. The Cochran’s Our study reveals that the sex instrumental heterogeneity has non-ignorable impact on sex-specific two-sample MR studies and the causal effects of anthropometric traits on breast/prostate cancer would be biased if sex-combined IVs are incorrectly employed. In the literature of causal inference in observational studies, Mendelian randomization (MR) is a novel statistical method to establish causal relationship between an exposure and an outcome by leveraging genetic variants as instrumental variables (IVs) . To guarHowever, due to the limitation of data sharing and participant privacy concern, individual-level GWAS datasets are often not accessible; instead, publicly available summary-level statistics are employed in practice, which brings one great benefit that the exposure and the outcome are not required to be measured on the same set of individuals, leading to the so-called two-sample MR study . Brieflyvia the two-sample MR. Here, we do not consider male breast cancer, as it only accounts for less than 1% of cases.Nevertheless, the two-sample MR still encounters many practical challenges that need to be addressed carefully. For example, the individuals of the two GWASs included in MR studies should take from non-overlapping populations; otherwise, misleading causal effect estimates may be generated . In addiHowever, this seems not to be true in sex-specific two-sample MR studies in terms of our literature review, and we discover that only a few MR studies mentioned in their analyses the problem of sex-specific instruments , 2. It iAlthough it is particularly important, a formal investigation about the sex instrumental heterogeneity in sex-specific two-sample MR studies is lacking and its consequence seems to be also overlooked by many MR researchers. As a result, improper causal inference might be generated . Therefo2 and study-specific covariates via linear regression. In addition, the SNP effect of WHR, WC, or HIP was estimated under the control of BMI. Based on these GWAS datasets, we yielded a set of uncorrelated associated SNPs (P < 5.00E-8) to serve as sex-combined or sex-specific IVs for each anthropometric trait , waist-to-hip ratio (WHR), waist circumference (WC), and hip circumference (HIP)] for individuals of European ancestry from the Genetic Investigation of ANthropometric Traits (GIANT) Consortium . For eacic trait .via logistic regression with principal components as covariates and sometimes additionally adjusted for study-specific covariates. The GWAS datasets employed in our study are summarized in We next acquired summary statistics of breast cancer from the Breast Cancer Association Consortium (BCAC)  and summg with all available SNPs (Z statistics of two traits. The regression slope provides an unbiased estimate for ρg. The software of LDSC (version v1.0.1) was downloaded from https://github.com/bulik/ldsc.To evaluate genetic similarity between the sex-combined and sex-specific effects of anthropometric traits as well as between the male-specific and female-specific effects of anthropometric traits, we applied cross-trait linkage disequilibrium score regression (LDSC) to calculate overall genetic correlation ρble SNPs . The LD ble SNPs  and thenrg among those independently associated SNPs that served as IVs. Note that, unlike ρg, which quantifies the global genetic overlap of two traits using genome-wide variants, rg can be viewed as a measurement of marginal genetic sharing between two traits because of only a small set of associated SNPs involved. We further assessed sex heterogeneity in each IV via the Cochran’s Q test. Specifically, the sex heterogeneity was tested based on sex-specific SNP effect estimates and standard errors; the P value of heterogeneity was corrected with the Bonferroni’s method to take multiple comparisons into account. Finally, the sex heterogeneity was quantified with the I2 statistic that was widely used in the literature.We also carried out the Pearson’s correlation analysis to quantify the genetic effect correlation m uncorrelated genetic variants with m following a uniform distribution ranging from 50 to 150. The minor allele frequency (MAF) of these SNPs was independently sampled from a uniform distribution ranging from 0.01 to 0.50. The genotypes (denoted by G1 or G2) were separately generated for N1 (1 × 105) male or N2 (1 × 105) female individuals under the assumption of linkage equilibrium and Hardy–Weinberg equilibrium (HWE). The effect sizes (denoted by α1 and α2) for the two sets of SNPs were drawn from a bivariate normal distribution with μ =  and Σ = , with rg varying from 0.1, 0.3, 0.5, to 0.7. Note that rg also partly quantified the genetic effect heterogeneity of the exposure between females and males for these SNPs; that is, smaller rg indicated larger heterogeneity. The residual error terms (denoted by e1 or e2) were separately sampled from independent standard normal distributions. We further rescaled α1 and α2 so that the phenotypic variance explained (PVE) by SNPs can be set to the given value for the male exposure and the female exposure.Given the potential sex instrumental heterogeneity in IVs , we implrg would not be impacted by the scaled effect sizes. After doing those, the exposures (denoted by x1 and x2) can be obtained.where δ is the scale parameter and can be estimated , while the sex-combined summary statistics of the exposure were yielded with the fixed-effects IVW meta-analysis based on the two sets of sex-specific summary statistics. The SNPs with marginal P values less than 0.05/m were identified to be female-specific or sex-combined IVs. If no SNPs satisfied this criterion, the one with the minimum P value would be employed. With the selected IVs, the female-specific and sex-combined causal effects of the exposure on the outcome were evaluated with the fixed-effects IVW MR approach  excluded SNPs that were not included in the breast/prostate cancer GWASs; (ii) excluded SNPs whose alleles were inconsistent between the exposure and the outcome; (iii) excluded SNPs that were likely related to breast/prostate cancer if the Bonferroni-corrected  studies . After tvia the IVW approach, we further implemented the weighted median method  by a simple u test:Finally, we formally compared the causal effects generated with sex-combined or sex-specific instruments  but less than 1 (H02: ρg = 1) (P = 0.076 for H02). In addition, the Pearson’s correlation analysis also showed that all the correlations between the sex-combined and sex-specific instruments are less than 1 (H30: rg = 1) (P = 0.472) for WC and P = 1.69E-5) for WHR between males and females  ; the onl rg = 1) . It need females , indicatQ test, we detect two out of 97 (2.1%) BMI-associated SNPs, four of 95 (4.2%) HIP-associated SNPs, 10 of 76 (13.2%) WC-associated SNPs, and 17 of 48 (35.4%) WHR-associated SNPs exhibit sex heterogeneity among candidate IVs  and all WHR-associated SNPs but three  are found to have larger effect sizes on females compared with males  (The results of simulation are displayed in  vs. 3%) . The mai vs. 3%) .rg = 0.1, 0.150 for rg = 0.3, 0.131 for rg = 0.5, or 0.127 for rg = 0.7. In contrast, approximate unbiased estimates of causal effects are generated when using sex-specific IVs. Overall, this simulation study explicitly reveals that the use of sex-combined IVs in sex-specific two-sample MR analyses may lead to a biased causal effect estimate in many cases and that the extension of the bias relies on the relative magnitude of the sex-specific effects and the sex-combined effects of the used IVs.Moreover, as can be expected, the sex-combined bias generally becomes larger as the sex heterogeneity in IVs increases . For example, when true effect size is 0.5 , the aveP = 0.003], between WC and breast cancer , as well as between BMI and prostate cancer  (Q test (P = 0.069), although other causal effects almost remain unchanged because only a few instruments are excluded, indicating that the sex heterogeneity in IVs might have a substantial influence on the statistical inference in sex-specific two-sample MR analyses.Like most previous studies , 2, we f= 0.022) , 4, indi= 0.022) . After t , WC is now only marginally associated with breast cancer (P = 0.057) (P = 0.060), although the causal effect is not influenced (0.865 vs. 0.862) (P = 1.63E-6), for HIP on breast cancer (P = 1.25E-20), as well as for WC on prostate cancer (P = 0.007) compared with those estimated with sex-combined instruments  . The ana. 0.862) . Third, truments , 4. Moretruments . For exatruments , 4, partGNAT2) has a large effect size of 0.071 on BMI and may be likely a potential outlier (P = 5.28E-04) (P = 0.418), ruling out the possibility of directional pleiotropic effects of instruments. The funnel plot also presents a symmetric pattern around the overall point estimate  , almost .28E-04) . The resestimate , implyinhttps://sb452.shinyapps.io/power  is slightly lower than that computed with sex-combined instruments (power = 98.3% when IVs = 66 and PVE = 1.42%) . The anaHowever, it is also shown that although there is a decrease in the number of valid IVs after using sex-specific instruments, the statistical powers of some associations are higher than those obtained with sex-combined ones , 4. For The main objective of our study was to investigate the influence of sex instrumental heterogeneity on causal effect estimation in sex-specific two-sample MR analyses where in principle sex-specific rather than sex-combined IVs should be employed. One of the common cases is the application of MR to breast cancer or prostate cancer, both of which lead to serious health threat in females and males worldwide . TherefoIn our simulation and empirical analyses, we illustrated that the sex instrumental heterogeneity had a non-ignorable impact on the causal inference and that the causal effects of anthropometric traits on breast/prostate cancer would be greatly influenced if sex-combined IVs were incorrectly applied. To the best of our knowledge, our study is among the first to formally examine the problem of sex-specific instruments in the two-sample MR.In sex-specific two-sample MR studies, the use of sex-combined instruments makes an implicit hypothesis that no effect differences exist between females and males, which, however, is not always satisfied in terms of previous observations e.g., . NeverthFrom a more generalized perspective, the females and the males can be viewed as two diverse populations that have different genetic foundations  as well Finally, we offer some suggestions for the issue of sex heterogeneity of instruments when conducting sex-specific two-sample MR studies. First, to guarantee to implement MR in the same population, it is necessary to check the original GWAS of exposures or contact authors to obtain sex-specific summary statistics; a clear statement about whether sex-specific IVs are employed is also highly recommended . Second,In summary, although the two-sample MR is technically easy to undertake, the principal modeling assumptions should still be validated in sex-specific two-sample MR studies strictly. Especially in the sex-specific two-sample MR analyses, the choice of appropriate IVs for exposures can reduce the bias of causal effect estimation and make MR results more reliable.Our study reveals that the sex instrumental heterogeneity may have a non-ignorable impact on sex-specific two-sample MR studies, and the causal effects of anthropometric traits on breast/prostate cancer would be biased if sex-combined IVs are incorrectly employed.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/PZ conceived the idea for the study. PZ, FG, and HZ obtained the data. PZ and YG cleared up the datasets, performed the data analyses, and drafted the manuscript. PZ, FG, HZ, YG, and JZ interpreted the results of the data analyses. All authors approved the manuscript and provided relevant suggestions.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Abstract. Acute tubular injury is the lesion most frequently described in this disease. However, four cases of ANCA-associated vasculitis (AAV) with COVID-19 with pauci-immune glomerulonephritis have recently been described. We report the case of an African woman, aged 70, in whom we diagnosed an AAV with pauci-immune glomerulonephritis in the context of COVID-19. She was treated with hydroxychloroquine and azithromycin for COVID-19. Corticosteroids and cyclophosphamide have been used for the treatment of vasculitis. The evolution was marked by the reappearance of COVID-19 one month after the beginning of an immunosuppressive therapy. The patient died a week later from respiratory failure. The occurrence of AAV during COVID-19 may not be due an unfortunate association but triggered by infection with SARS-CoV-2. The use of immunosuppressive therapy should be discussed due to the potential risk of reactivation or recurrence of the viral infection.  Coronavirus disease 2019 (COVID-19) is a new disease that appeared at the end of 2019. It is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 is mainly characterized by respiratory manifestations but can affect other organs, in particular the kidneys. However, not every kidney damage due to this disease is fully understood. Acute tubular injury (ATI) and collapsing glomerulopathy are the lesions most frequently described in this disease . There a3, the C-reactive protein was elevated to 260 mg/L. Serum creatinine was normal on admission at 12 mg/L, and blood urea was 0.36 g/L. The diagnosis of COVID-19 was retained by a positive SARS-CoV-2 reverse transcription polymerase chain reaction (RT-PCR). The chest CT scan performed without injection of contrast medium showed ground-glass opacities in both pulmonary fields. The patient was put on hydroxychloroquine (HCQ) 600 mg/day, azythromycin (AZT) 500 mg/day, and dexamethasone 6 mg/day. During hospitalization, the patient presented an acute kidney injury (AKI) with serum creatinine at 24 mg/L then at 36 mg/L. Proteinuria was 0.6 g/day, and urine sediment was active with hematuria at 20 red blood cells per high-power field (RBC/hpf) with hematuria with blood cylinders. Two weeks after diagnosis, the patient was cured of COVID-19 with two negative control PCR tests. The follow-up chest CT scan was normal. However, renal failure persisted with worsening renal impairment. The kinetics of serum creatinine are shown in We describe a 70-year-old African woman with history of hysterectomy for uterine myomatosis 10 years ago. She was seen in the emergency room with complaints of fever, cough, and rhinorrhea. On physical examination, blood pressure was 120/70 mmHg, heart rate 112 beats/min, temperature 40 °C, respiratory rate 24 cycles/min, and weight 82 kg. The blood count showed a hemoglobin level of 11.2 g/dL, hyperleukocytosis at 19,000/mmCurrently, it is clearly established that some forms of vasculitis may be secondary to viral infections. This is the case with hepatitis B virus-related polyarteritis nodosa and hepatitis C virus-related cryoglobulinemic vasculitis. For SARS-CoV-2, although the association with vasculitis has not yet been confirmed, the onset of cases of Kawasaki disease related to SARS-CoV-2 infection is an example of SARS-CoV-2 triggering vasculitis . If we lIn terms of clinical and biological manifestations, there is no difference between our patient and four other cases already described. We observed that the association of COVID-19 and AAV can occur regardless of age or gender. Our patient did not have extrarenal clinical manifestations of vasculitis. When we reviewed the cases already described in the literature, only one presented an extrarenal sign such as arthritis . Intra-aThe treatment remains the conventional treatment of AAV, based on corticosteroids associated with an immunosuppressant, either CYC or rituximab (RTX) . This diHerpesviridae. Immunosuppressive treatment could lead to reactivation of SARS-CoV-2, which would have remained in the latent phase. However, it should be noted that in the four published cases of AAV and COVID-19, a reappearance of COVID-19 was not noted in any patient. The reappearance of COVID-19 in our patient raises two questions. First, we can think of a mutation of the virus, a reinfection by another strain of the SARS-CoV-2. Second, we can ask whether the virus persisted in the body after recovery from the disease. That is, the virus remained in the body in a latent state as in some viral infections such as Epstein-Barr virus infection and infection with The occurrence of AAV during COVID-19 may not be a random association but triggered by infection with SARS-CoV-2. The use of immunosuppressive therapy should be discussed because of the potential risk of reactivation of the viral infection. The present case report adhered to the Declaration of Helsinki. Written informed consent for publication was obtained from the patient’s son. This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors. The authors have no conflict of interest to declare.


Here, we present the synthesis and bulk co-assembly of pyrene (Pyr) and naphthalenediimide (NDI) functionalized oligodimethylsiloxanes (oDMS) of discrete length. We tune the donor–acceptor interactions by connecting the pyrene and NDI to the same oligomer, forming a heterotelechelic block molecule (NDI-oDMSPyr), and to two separate oligomers, giving Pyr and NDI homotelechelic block molecules (Pyr-oDMS and NDI-oDMS). Liquid crystalline materials are obtained for binary mixtures of Pyr-oDMS and NDI-oDMS, while crystallization of the CT dimers occurred for the heterotelechelic NDI-oDMS-Pyr block molecule. The synergy between crystallization and phase-segregation coupled with the discrete length of the oDMS units allows for perfect order and sharp interfaces between the insulating siloxane and CT layers composed of crystalline CT dimers. We were able to tune the lamellar domain spacing and donor–acceptor CT interactions by applying pressures up to 6 GPa on the material, making the system promising for soft-material nanotechnologies. These results demonstrate the importance of the molecular design to tune the CT interactions and stability of a CT material.The assembly of donor–acceptor molecules  A robust, soft material held together by charge transfer interactions is developed. The nanostructure dimensions and material properties can be tuned by molecular design or pressure. Although donor–acceptor crystals are known for decades, the perfect arrangement is not achieved in soft (block copolymer) materials yet due to the disorder in polymeric materials. Here we introduce the concept of block molecules that are in-between crystals and polymers. They combine the perfect 2D crystallisation of lamellae while they still have the disordered part connecting the 2D crystalline parts. Pressure-induced tuning of the charge-transfer distance gives new directions for novel materials.1–5 The well-known examples are the many studies by Seth Marder and his group on the use of CT interactions in functional materials. His originality in bringing new concepts to materials science has inspired many of us.The alternate stacking of donor–acceptor (D–A) molecules driven by charge-transfer (CT) interactions has resulted in the development of various materials with fascinating electronic properties.6 Alternatively, conductivity in CT cocrystals may be introduced upon applying pressure, inducing a phase-transition.7 Pressure-induced structural changes in these CT crystals result in a change of resistivity and thereby these systems become semiconducting, or even superconducting.8–10 For these electronic properties, the alternate stacking of donor and aceptor molecules is crucial and therefore the functional CT materials mostly comprise CT cocrystals.11,12A CT cocrystal that exhibits ferroelectricity at room temperature was reported by Stupp and Stoddart and coworkers.13,14 liquid crystals,15,16 and crosslinked supramolecular networks,17,18 have gained increasing attention recently.19 These materials are composed of hierarchical structures that are micelles, nanotubes or fibers, assembled via CT complexation of π donors and acceptors. In contrast to hydrogen-bonded supramolecular assemblies,20 supramolecular stacks of donor–acceptor molecules are less extensively explored both in bulk and solution due to their comparatively low association constant.19 This typically results in self-sorting, which limits the formation of organized, multicomponent nanostructures held together by intermolecular D–A complexation.21CT soft materials, such as gels,22–24 metal ion complexation,25 peptide-mediated assembly26,27 or amphiphilicity.28,29 Moreover, a geometrical fit between D and A improves the association since π-stacking is maximized by an increased contact area between the building blocks.30,31 Therefore, the acceptor naphthalene diimide (NDI) is often combined with the donor pyrene (Pyr). In crystals, this combination results in D–A CT dimers that stack well in the unit cell,32,33 while in polymeric and soft materials, the co-assembled packing of Pyr and NDI is much less ordered.34To arrive at hetero-aggregation between the D–A molecules, additional, supramolecular interactions are often included in the molecular design. Examples involve hydrogen-bonding,35 Over the past decades, the covalent attachment of siloxanes to NDIs or pyrenes was studied thouroughly to obtain phase-segregated assemblies for e.g. self-healable and adaptive materials,36–38 or flexible devices.39–41 The highest degree of order is obtained for small molecule siloxane conjugates due to their discrete length. Recently, some of us pushed the boundaries of NDI–siloxane assemblies to a length scale between polymers and small molecules using discrete length oligodimethylsiloxanes (oDMS) end-capped with NDIs.42 The incompatibility of the oDMS and NDI parts in combination with crystallization of the NDIs yielded highly ordered lamellar morphologies with sub-10 nm domain sizes. The NDI–siloxane conjugates were utilized to pattern a graphite surface.43 Moreover, we have reported previously that conjugation of small molecules to discrete siloxanes can result in order over macroscopic length scales,44,45 giving unique properties to these siloxane-based materials.46Nanoscale order has been achieved in supramolecular polymers by phase segregation.Here, we report on the co-assembly of discrete NDI- and Pyr-oDMS block molecules to study the CT complexation of NDIs and pyrenes assisted by phase-segregation in bulk materials . As a re8-NDI. The pyrene-functionalized siloxanes were synthesized from 1-hydroxypyrene (1) and 5-bromo-1-pentene (2), forming the olefin-functionalized pyrene 3, which was used as the common building block for all Pyr-oDMS conjugates , respectively. NDI-1 was obtained from naphthalene tetracarboxylic dianhydride (4), n-pentylamine (5) and 4-penten-1-amine using a sequential microwave-assisted protocol (51%)  with the pyrene on one chain-end and the NDI on the other end was synthesized from pyrene 3 and NDI 7. For this, a Karstedt catalysed hydrosilylation reaction was performed with one equivalent of the pyrene precursor and two equivalents of oDMS8 dihydride to prefer the formation of monofunctionalized siloxane (8-NDI (45%). All end-functionalized siloxanes were obtained in high purity and characterized as presented in the ESIWe synthesized homotelechelic oligomers Pyr-1, Pyr-2, Pyr-3, and NDI-1, as well as a heterotelechelic oligomer denoted as Pyr-Sinjugates . The oDMol (51%) .42 Finalsiloxane . However−1) to room temperature. We here show the bulk co-assembly of Pyr-1 with NDI-1, denoted as Pyr-1:NDI-1 , followed by evaporation of the solvent. To exclude any solvent effects the material was molten at 120 °C and slowly cooled  shows a cold crystallization temperature (Tcc) followed by a melt transition temperature (Tm) and a broad endothermic transition upon heating (TODT) temperature as the enthalpic energy of the transition is relatively low (<3.6 kJ mol−1) (TDOT) is observed at 84.1 °C, followed by a crystallization transition below room temperature (−1.7 °C). These results indicate the presence of a liquid crystalline phase at room temperature which is in accordance to the waxy appearance of the material. Moreover, birefringent textures were observed by polarized optical microscopy (POM) indicative of liquid crystalline ordering at room temperature  , entry 4q* and its integer multiples (q2*) next to the primary scattering peak q* is observed. The shoulder completely vanished at 60 °C upon heating, while it reappeared at 20 °C upon cooling and became even more pronounced at −20 °C  and Tc (−1.7 °C).We investigated the liquid crystalline order in the Pyr-1:NDI-1 mixture by medium- and wide-angle X-ray scattering (MAXS and WAXS) . The extultiples . In the q region (q > 7 nm−1) for Pyr-1 and NDI-1, indicative of a highly crystalline structure  at 67.3 °C with a relatively small enthalpic contribution (2.2 kJ mol−1), indicative of an order–order transition. This was followed by melting of the material at 92.2 °C (Tm) with an energy release of 10.9 kJ mol−1. Subsequent cooling gave a crystallization transition temperature (Tc) at 87.6 °C, followed by a weaker exothermic transition at 48.4 °C, which is assigned to an order–order transition (T2). The formation of small, birefringent spherulites was observed under the polarized optical microscope when a film of Pyr-Si8-NDI was cooled from the isotropic melt to 80 °C between two glass plates  and 19.1 nm−1 (π2), representing distances of 0.35 and 0.33 nm, respectively. We attribute the distances of 0.33 (π2) and 0.35 (π1) nm to the π-stacking distance of a pyrene and NDI within a CT dimer and the distance between CT dimers, respectively.32 Therefore, we suggest that the pyrenes and NDIs form alternating stacks at room temperature, as schematically illustrated in 32 but has not been observed for soft, polymeric or oligomeric CT materials to date. We expect that the high, crystalline order is due to the discrete design of the block molecules allowing for a perfect, defect free packing of the NDIs and pyrenes.The 1D transmission scattering profile of the Pyr-Sies of q* . This in1 and π2 scattering peaks merged into one peak at 80 °C in the variable temperature transmission scattering data . As a result, the intensity of the CT band in the absorption spectrum strongly decreases, although the band is still present  analysis  – perpendicular to the lamellar domain spacing – also decrease upon applying pressure .The change in optical properties of Pyr-Si 6.3 GPa . The pea 6.3 GPa . The CT  6.3 GPa . The peaECT) (2-stacking distance (1/dπ2)   and plot (1/dπ2) . We findECT = E+D/D − E−A/A + ECoul with E+D/D the oxidation potential of the donor, E−A/A the reduction potential of the acceptor and ECoul a term representing the coulombic electrostatic potential energy between the ionized donor D+ and acceptor A− in the CT state. In the simplest possible description, one treats the ionized donor and acceptor as point charges so that ECoul = −qe2/4πε0εrdπ2 with qe the electron charge, ε0 the permittivity of vacuum, εr the relative dielectric constant and dπ2 the closest distance between donor and acceptor. If we assume E+D/D and E−A/A to be independent of applied pressure then the above expression for Ecoul indeed accounts for the correlation observed in ECT state and 1/dπ2. Moreover, assuming a value εr = 2 similar to benzene, one predicts a slope −qe2/4πε0εr = −7.2 (eV Å) for the correlation between ECT and 1/dπ2, while from a fit to the experimental data in ECTvs. 1/dπ2 supports our hypothesis that the redshift of fluorescence from the material at increased pressure is due to a reduction in the distance between Pyr and NDI moieties under compression.We hypothesized that the red shift in charge transfer luminescence with increasing pressure is due to the closer proximity of the NDI and pyrene cores. The energy of the CT state may be approximated as 8-NDI with anisotropic lamellar domains are promising, allowing the tuning of the properties and distances in one direction instead of the average. In particular, lamellar domains that are oriented perpendicular to the pressing direction are highly interesting, as the Pyr and NDI molecules could possibly be pushed closer to each other and thereby lower the energy of the CT state even further.In the above-described experiments, the pressure is applied on one side of the material and therefore we have to consider the direction of the lamellae in the material. The material is composed of isotropic ordered lamellar domains and therefore pressing in one direction of the material gives an average of the compression result. Pressing perpendicular to the lamellae affects mostly the distance between Pyr and NDI, while the siloxane is not much affected by the pressure in this direction. In contrast, pressing parallel to the lamellae compresses the siloxane significantly and the Pyr and NDI cores are not pushed together. As the material is composed of isotropically ordered lamellae, the lamellae exist in all angles towards the applied pressure direction. Therefore, average change in domain spacing, π-stacking distance, compressibility and fluorescence is obtained. Hence, future studies on Pyr-Si8-NDI and two-component Pyr-1:NDI-1 show striking differences. Therefore, we compare Pyr-Si8-NDI with Pyr-1:NDI-1, which are similar in terms of alkyl and siloxane linker lengths and stoichiometry of the D–A molecules, only the connectivity differs. In both cases, a lamellar nanostructure with a domain spacing of 3.1 nm was obtained. Nevertheless, two major differences in the CT material properties and nanostructure of the one-component Pyr-Si8-NDIversus the two-component Pyr-1:NDI-1 assemblies were observed. First, the highly efficient packing of the NDIs and pyrenes in the one-component system resulted in the formation of perfectly packed CT dimers at room temperature, while a more random distribution of pyrenes and NDIs within the stacks is observed for the two-component mixture. For the one-component Pyr-Si8-NDI system, we distinguish between the π-stacking distance of the CT dimers and within the dimer concluding a crystalline, alternate stacking of the pyrenes and NDIs which has not been reported for soft CT materials to date. In general, soft, non-crystalline CT materials show a single, broader π-stacking scattering peak at 0.34 nm,34 similar to the two-component Pyr-1:NDI-1 mixture at room temperature and Pyr-Si8-NDI above T1. The single π-stacking scattering peak indicates a random distribution of the NDIs and pyrenes in the CT stacks. Second, the CT nanostructure of Pyr-Si8-NDI is thermodynamically stable at room temperature as its order–order transition takes place at 48.8 °C. In contrast, the order–order transition of Pyr-1:NDI-1 is below room temperature (−1.7 °C). Hence, the two-component material is liquid crystalline at room temperature and therefore more fluctuations in the nanostructure are possible, while the one-component material is highly crystalline at room temperature.The assembly morphology, CT properties, and stability of the CT state in the one-component Pyr-Si8-NDI system is able to endure high pressures while the two-component materials are waxes and therefore not suitable for pressure experiments. Nevertheless, the nanostructures of the two-component materials are readily changed by varying the length of one of the two components.Thereby, the Pyr-Si31 – impossible in the bulk material. In particular, a pyrene in Pyr-Si8-NDI may interact with the NDI on the other end of the siloxane linker or with an NDI of another Pyr-Si8-NDI oligomer. We speculate that the difference in properties and packing between Pyr-Si8-NDI and Pyr-1:NDI-1 could be due to a larger number of assembly possibilities for the two-component Pyr-1:NDI-1 system which is a well-known phenomenon in supramolecular copolymers in solution.51,52 This makes the formation of a thermodynamically stable CT structure for Pyr-1:NDI-1 more demanding than for the heterotelechelic Pyr-Si8-NDI one-component system. Alternatively, the differences in CT properties for the one- and two-component assembly may arise from the highly effective molarity, potentially accessible with Pyr-Si8-NDI, as already shown in other supramolecular systems.53 Indeed, in solution we could observe this effect showing a clear CT band at 550 nm in the absorption spectrum of Pyr-Si8-NDI in methylcyclohexane (MCH), while no CT band was observed for the Pyr-1:NDI-1 mixture in MCH  and naphthalenediimide (NDI) peripheral blocks. We have shown that the covalent attachment of Pyr and NDI moieties by a siloxane linker in a heterotelechelic design influences the material properties and nanostructure significantly when compared to a homotelechelic, binary mixture. The co-assembly of homotelechelic Pyr- and NDI-oDMS resulted in a liquid crystalline material with a lamellar nanostructure in which the NDIs and pyrenes are randomly distributed throughout the Pyr/NDI layer. A highly ordered and thermodynamically stable CT material was formed by the heterotelechelic siloxane resulting from phase segregation induced by the siloxane, in synergy with co-crystallization of the NDI and pyrene. Herein, the NDIs and pyrenes form stacks characterized by crystalline CT dimers featuring an alternation of NDI and Pyr units. These are isolated in layers by the amorphous siloxane, forming a highly ordered lamellar nanostructure. The crystalline material is very robust and can undergo pressures up to 6.1 GPa at room temperature without losing the morphology. The NDI- and Pyr-cores are brought in closer proximity by pressure, resulting in charge delocalization which makes this Pyr-SiAll together, we have shown the importance of the molecular design on the nanostructure and CT properties in a multicomponent, bulk assembly system. We were able to tune the Pyr–NDI interactions and nanostructure feature sizes by the molecular design and upon applying pressure. Thereby, we created a modular and robust system in which the distance between the CT layers and between the donor and acceptor molecules can be altered making these materials promising for soft nano-electronics.B. A. G. L., G. V., A. R. A. P., and E. W. M. conceived the project and directed the research. B. A. G. L., B. F. M. d. W., J. A. B., and F. V. d. G. conducted the synthesis, B. A. G. L., M. H. C. v. S., B. W. L. v. d. B. and F. V. d. G. characterized the morphologies, S. C. J. M. analyzed the optical measurements, K. K., H. S., and T. A. performed the high-pressure experiments, B. A. G. L. and E. W. M. wrote most of the paper and all authors contributed to the writing and editing of the final manuscript.There are no conflicts to declare.MH-009-D1MH01141C-s001


Furthermore, the PANI-MWCNT-APTES-modified GCE current response was stable, repeatable, reproducible, and short. In addition, PANI-MWCNT-APTES/GCE was harnessed for the first time for cadmium detection in real water samples, and the result was satisfactory. Therefore, the recorded results suggest that the newly designed PANI-MWCNT-APTES is a promising material for detecting Cd in the near future for human health and environmental protection.Cadmium belongs to the group of potentially toxic metals that have high health and environmental significance. Due to its adverse effects on the environment, this study develops an effective electrochemical sensor for detecting a polyaniline-multiwalled carbon nanotube-3-aminopropyltriethoxysilane (PANI-MWCNT-APTES) substrate cast on the GCE. The as-prepared PANI-MWCNT-APTES was prepared by a wet chemical method, and its formation was investigated using several techniques. As a result, the prepared material exhibited a limit of detection of 0.015 µM for cadmium ions (Cd This ranged from 0.05 µM to 50 µM. However, the FDR indicates the full concentration range at which current response to the change in concentration was noticed. For this study, this ranged from 0 to 50 µM.2+ ions was obtained using the following formula:The sensitivity of PANI-MWNT-APTES-modified GCE to CdSensitivity = slope of the calibration/surface area of the GCE−1 cm−2, as shown by The obtained value was 4.237 µA µMResponse time is a significant parameter in sensing applications because it indicates the time frame for the electrochemical process. Usually, a sensor with a fast response time is desirable for in situ analysis . For thiThe stability of the current response of PANI-MWCNT-APTES was investigated by repetitive measurement techniques (repeatability and reproducibility). While repeatability indicates the accuracy of a method, reproducibility shows the precision of the sensor’s performance. For this study, ten consecutive measurements at the same experimental condition were conducted, and the obtained result is given in 2+ has two positive charges (2+) and is prone to interferents from metal ions with similar charges. Therefore, likely interfering ions mostly considered for this study are Mg2+, Mn2+, Ni2+, Ca2+, Zn2+, Yt3+, NO2−, and PO43−. The obtained result is given in 2+ ions was reduced slightly. However, the relative standard deviation (RSD) of the mean value of Cd2+ ions is less than 5%. Therefore, the response of PANI-MWCNT-APTES is stable in the matrix containing likely interferents.Usually, metal ions with similar charges tend to interfere with the redox reaction of each other. For instance, CdAs indicated in 2+ ranged from 103.6% to 129.9%. Thus, the obtained result is satisfactory and suitable for use in real water samples.The percentage recovery for the spiked Cd2+ using different methods. Compared with the earlier reported method, the current study compares very well with the literature and even outperformed many. Significant progress on Cd2+ ion assays is the stability of PANI-MWCNT-APTES-modified GCE towards the Cd2+ reduction process. The typical setback in metal ion detection is electrode fouling, which makes the substrate  on the GCE have an unstable current response. However, it is suggested that crosslinking of MWCNT to the PANI-APTES matrix could have contributed to the anti-fouling nature of the PANI-MWCNT-APTES substrate.2+ ions in water using a newly synthesized composite based on PANI-MWCNT-APTES. The composite was synthesized by the wet chemical method, and its successful synthesis, including XRD, XPS, FESEM, TGA-DTA, FTIR, and CV. The synthesized PANI-MWCNT-APTES was applied for electrochemical sensing of Cd using linear sweep voltammetry. The sensor gave a linear response to Cd2+ ions in the range of 0.05 µM to 50 µM with a limit of detection of 15.5 nM. It also displayed high sensitivity and stability. In addition, the developed sensor based on a PANI-MWCNT-APTES composite exhibited fast response time, and its current responses were not affected by likely interferents. It was also used satisfactorily for Cd2+ ion determination in real water samples. Therefore, this study provides a cheap and effective method of Cd2+ ion detection in the environment for human health safety and environmental pollution prevention.The current study focused on detecting Cd


Glycosylphosphatidylinositol (GPI) anchor modification is a posttranslational modification of proteins that has been conserved in eukaryotes. The biosynthesis and transfer of GPI to proteins are carried out in the endoplasmic reticulum. Attachment of GPI to proteins is mediated by the GPI-transamidase (GPI-TA) complex, which recognizes and cleaves the C-terminal GPI attachment signal of precursor proteins. Then, GPI is transferred to the newly exposed C-terminus of the proteins. GPI-TA consists of five subunits: PIGK, GPAA1, PIGT, PIGS, and PIGU, and the absence of any subunit leads to the loss of activity. Here, we analyzed functionally important residues of the five subunits of GPI-TA by comparing conserved sequences among homologous proteins. In addition, we optimized the purification method for analyzing the structure of GPI-TA. Using purified GPI-TA, preliminary single particle images were obtained. Our results provide guidance for the structural and functional analysis of GPI-TA. Protein anchoring by glycosylphosphatidylinositol (GPI) is a conserved posttranslational modification in eukaryotes ,2,3,4. MGPI biosynthesis and transfer to proteins are carried out on the endoplasmic reticulum (ER) membrane. The synthesis of GPI is started from phosphatidylinositol (PI) by the stepwise addition of sugars, a fatty acid, and ethanolamine-phosphates (EtNPs). GPI precursor proteins, which have an N-terminal signal sequence for ER translocation and a GPI attachment signal peptide at the C-terminus, are separately synthesized . Once the proteins are translocated into the luminal side of the ER, the N-terminal signal sequence is cleaved by signal peptidases, and the C-terminal GPI attachment signal peptide is recognized by GPI-transamidase (GPI-TA), which transfers a GPI to the proteins .Mammalian GPI-TA is a multi-subunit complex consisting of five subunits: PIGK, GPAA1, PIGT, PIGS, and PIGU . PIGK is the catalytic subunit of GPI-TA, and is homologous with members of the cysteine protease family with transaminase activity . PIGK liPIGK . StrepTactin Sepharose was collected through the column and washed 10 times with 5 mL of buffer A [W]. The GPI-TA complex was eluted by 5 mL of buffer B ) five times .All the purification steps were performed at 4 °C. K562K-hPIGK-StrepII cells (~50 L) were harvested by centrifugation and washed with PBS. The cells were disrupted with a microfluidizer in TBS  , and the debris was removed by centrifugation  was used according to the instructions. The protein bands were cut, and the peptides were analyzed using mass spectrometry to verify whether GPI-TA was successfully purified.− s−1, and each movie was 7.6 s long, dose-fractioned into 38 frames with an exposure of 1.18 e− Å−2 for each frame.A total of 2.5 μL of the GPI-TA complex sample was applied to a glow-discharged holey carbon-film grid  blotted with a Vitrobot system  using a 3.0 s blotting time with 100% humidity at 9 °C, then the sample was plunge-frozen in liquid ethane. Cryo-EM data collection was performed using a 300 kV Titan Krios microscope (FEI) equipped with a K2 summit direct electron detector camera  set to super resolution mode, with a pixel size of 0.65 Å  and a defocus ranging from −1.5 µm to −2.3 µm. The specimen stage temperature was maintained at 80 K. The dose rate was 10 eA total of 2445 movies were motion-corrected and binned with MotionCor2  with 5 ×


To evaluate the effects of anti-VEGF treatment of neovascular age-related macular degeneration (nAMD) in a real-life clinical setting.  Study design is a retrospective case series. Naïve nAMD patients treated with intravitreal injection of aflibercept or ranibizumab were analyzed over a 24-month follow-up. Each patient received the loading dose, followed by a PRN regimen. Patients were further subdivided into subgroups according to macular neovascularization type, best corrected visual acuity (BCVA) at baseline (BCVA > 0.3 LogMAR and BCVA ≤ 0.3 LogMAR), and different anti-VEGF drugs. Primary outcome was the changes in BCVA and central macular thickness (CMT) over 24 months. Secondary outcomes included the influence of the selected drug and of the baseline BCVA on the final outcomes. p < 0.01). Classic, polypoidal choroidal vasculopathy and mixed subtypes significantly correlated with worse visual outcome (p < 0.01). Overall, baseline BCVA significantly correlated with both 1-year and 2-year follow-up changes (p < 0.01). Moreover, BCVA at 1-year significantly correlated with BCVA changes at 2-year follow-up (p < 0.01). Furthermore, CMT changes from baseline significantly correlated with both 1-year and 2-year follow-up measurements (p < 0.01).  439 patients  with naïve AMD-related macular neovascularization were included in the analyses. Mean age was 78 ± 8 years old. Compared to baseline evaluations, not significant BCVA changes were found at 1-year and 2-year examinations. CMT was significantly reduced at both 1-year and 2-year follow-ups ( Anti-VEGF approach is generally effective in stopping nAMD progression in our real-life analysis. No difference was found comparing patients treated with ranibizumab and aflibercept, nor in patients with drug switching. Age-related macular degeneration (AMD) is a leading cause of vision loss which affects people older than 50 years old in developed countries. The European prevalence in 2040 is estimated to reach 21 million of people affected by the early stage and the 4.8 million by the late one –6. NeovaThe study was designed as a retrospective investigation to describe the real-life management in patient affected by nAMD treated with ranibizumab or aflibercept over a 24-month follow-up. Switching to a different anti-VEGFdrug was allowed at ophthalmologist's discretion on the basis of predefined criteria. Study protocol was approved by the Institutional Review Board of Scientific Institute San Raffaele, and the procedures followed the straits of the Declaration of Helsinki. Written informed consent was acquired from all the patients included in the present study. Patient's recruitment started from January 2014 and ended in December 2016 in order to permit the follow-up analysis.Naive neovascular AMD was diagnosed on the basis of clinical data and multimodal imaging. Patients underwent a loading dose of 3 injections, followed by a PRN treatment regimen with monthly examination. Retreatments were administered when reactivation or progression of the disease was registered, on the basis of the identification of retinal or subretinal fluid on optical coherence tomography (OCT), leakage on fluorescein angiography/indocyanine green angiography, or new macular hemorrhages.μm intervals, covering an area of 20 × 15 degrees (approximately 6 × 4.5 mm2).Neovascular AMD was diagnosed through dilated fundoscopy examination, by OCT, and fluorescein angiography. Patients underwent monthly ophthalmological examinations, including measurement of best corrected visual acuity (BCVA), using standard ETDRS charts, funduscopicexamination, Goldman applanation tonometry, and OCT . The acquisition protocol used a 19-line raster SD-OCT scheme, with each line at 240 Any other retinal disorders or any condition (systemic diseases or treatment) which could interfere with the anti-VEGF therapy or the clinical outcomes was considered as exclusion criteria.The criteria followed to the treatment switching included CMT reduction < 30% and/or BCVA deterioration > 10 ETDRS letters.Primary outcomes were the change in BCVA and central macular thickness (CMT) over the 24-month follow-up. Secondary outcomes included the influence of the selected drug and of the baseline BCVA on the final outcomes.p value <0.05 is estimated as statistically significant. Comparison between parameters at baseline and follow-ups were expressed by means of coupled t-test.Statistical analysis was achieved by both Microsoft Excel 2013  and SPSS Statistics Version 23.0 . Each result is expressed as mean ± SD (standard deviation), and a In order to analyze subgroups of patients, the analysis of variance (ANOVA) followed by the Tukey HSD test for posthoc analysis was used.Correlation analysis between numeric variables was obtained and analyzed with Tau-Kendall correlation coefficient.p > 0.05) that were evaluated. The entire cohort was subdivided in different groups using a BCVA cut-off (0.3 LogMAR) and MNV types. Percentages for each CNV type were 56.8% for type 1 (occult form), 26.1% for the classic form (type 2), 13.5% for mixed form, and finally, 3.6% for polypoidal choroidal vasculopathy. Ranibizumab or aflibercept monotherapy was used in most of the cases over the follow-up, whereas a shifting was registered in 28% of cases that was also made.Overall, 479 naïve AMD-related macular neovascularization (MNV) patients were recruited. Twenty patients were excluded due to high media opacity; another 20 patients were excluded because of uncontrolled arterial hypertension (12 patients) and glaucoma (8 patients). The mean age was 78 ± 8 years old; 51% were male, and the 49% were female . On the other hand, CMT was significantly reduced at both 1-year and 2-year follow-ups   .p < 0.01)  .p < 0.01) (p < 0.01)  . In part < 0.01) . Further < 0.01) .p < 0.01) compared to patients with better baseline BCVA (≤0.3 LogMAR), at both baseline (416 ± 136 vs 343 ± 83 μm) and 1-year (359 ± 105 vs 308 ± 71 μm) measurements (Eyes with worse baseline BCVA (>0.3 LogMAR) were characterized by greater mean CMT (urements . Patientp < 0.01), without remarkable functional and anatomical changes , at the end of the follow-up, their CMT turned out to be similar (314 μm vs 300 μm). This may be explained as the consequence of adopting CMT as a clinical parameter to guide the retreatment planning.Our study described the achieved outcome of nAMD patients treated by anti-VEGF injections in real-life. Despite the positive results, application of clinical trial strategy in the real-world practice is hardly achievable because of many reasons, especially regarding the strict inclusion and exclusion criteria along with the patients' compliance. In real-life setting, missed appointments, which elongate intervals between injections and also comorbidities, which discourage to attend visits, are frequent . For theOur results confirm that the anti-VEGF approach in the real-life practice can lead to a stabilization of the visual function, stopping the natural progression towards a visual loss. In particular, our data are in line with those of the COMPASS study .On the oAlmost one third of our patients were shifted from one drug to another one on the basis of the unsatisfactory response at the ophthalmologist's discretion. Nevertheless, no improvement in BCVA and CMT was registered in the patients shifted to the new agent, even though the global number of injections increased because most of the ophthalmologist decided to restart with a loading phase on the new drug. Contrasting results have been obtained by other studies –18, but MNV type stratification found that, differently from classic, polypoidal choroidal vasculopathy and mixed pattern, occult MNV (36% of the cohort) showed better outcomes comparing baseline to year 1, baseline to year 2, and even year 1 to year 2.Overall, our retrospective analysis shows a clinical outcome slightly inferior to those revealed by ANCHOR and MARINA trials , 20.The More specifically, the most important causes included the distance of the patient from the hospital, comorbidities that can discourage patients to be treated, and long waiting lists, which elongated the intervals among injections. In addition, we need to underline that the retrospective design of the study is an inherent bias per se. Moreover, we are aware that the adopted PRN treatment regimen may have an impact on the outcome achieved, since PRN methodology is known to yield inferior outcome, compared with fixed or treat and extend regimens. Moreover, we are aware that the lack of extremely positive outcome may be explained considering that real-life setting is quite different from sponsored studies.Further studies on national basis are warranted to implement the real-life use of the anti-VEGF approach for the management of AMD.


A rapid, sensitive and simple microflow cytometry-based agglutination immunoassay (MCIA) was developed for point-of-care (POC) quantitative detection of SARS-CoV-2 IgM and IgG antibodies. The antibody concentration was determined by using the transit time of beads aggregates. A linear relationship was established between the average transit time and the concentration of SARS-CoV-2 IgM and IgG, respectively. The limit of detection (LOD) of SARS-CoV-2 IgM and IgG by the MCIA measurement are 0.06 mg/L and 0.10 mg/L, respectively. The 10 µL sample consumption, 30 min assay time and the compact setup make this technique suitable for POC quantitative detection of SARS-CoV-2 antibodies.  Despite the COVID-19 pandemic persisting for more than a year, there is still a need for a fast and sensitive antibody test to detect SARS-CoV-2 infections. Unlike diagnostic tests, antibody or serological tests are not intended to detect infections at their onset. Rather, they detect if an individual has produced antibodies in response to a specific foreign body, in this case, the SARS-CoV-2 virus. Since seroconversion occurs some time after initial infection, antibody tests are not intended to replace diagnostic testing methods, namely PCR tests [POC antibody tests are serological tests that do not require the use of laboratory technicians or facilities to provide results . POC tesCurrently, enzyme-linked immunosorbent assays (ELISAs) and lateral flow immunoassays (LFIAs) are two common methods used to detect SARS-CoV-2 antibodies. ELISAs must be performed by skilled operators, have long incubation times and require laboratory equipment to quantify results, making them impractical for POC environments . LFIAs, Other testing formats have been proposed for the detection of SARS-CoV-2 antibodies including flow cytometry. Grzelak et al. developed a technique to detect anti-spike (S) protein antibodies by expressing the S protein on the surface of human embryonic kidney 293T cells. The cells were then incubated with human serum samples and fluorescent anti-IgG or anti-IgM before being evaluated by flow cytometry . HorndleIn this work, a fast and sensitive MCIA suitable for the POC quantitative detection of SARS-CoV-2 IgM and IgG was proposed. Microbeads were conjugated to anti-IgM and anti-IgG antibodies and incubated with SARS-CoV-2 antibodies. SARS-CoV-2 S protein was then added to cause microbeads to agglutinate. Using microflow cytometry, transit time was measured and used to distinguish monomers from larger aggregates. The relationship between the average transit-time of the aggregates and certain concentrations of SARS-CoV-2 IgM and IgG was studied. A linear correlation was found. This work also discussed the potential application of the MCIA for the POC detection and quantification of SARS-CoV-2 IgM and IgG.7 beads/mg. The MBs need to be washed before the antibody coupling procedure. A total of 5 mg of lyophilized beads were weighed out and resuspended in 1 mL of 0.1 M sodium phosphate buffer (PB) . The MBs were incubated for 10 min with tilting and rotation and then applied to a magnet. The supernatant was removed by a pipette. The MBs were suspended in 1 mL of PB again and vortexed for 30 s. The MBs were ready to couple to secondary antibodies after removing the supernatant. A total of 5 mg of MBs were used to couple approximately 100 µg of secondary antibody as recommended by the user manual. The concentration of goat anti-human IgM secondary antibodies was 2.32 mg/mL. Therefore, the volume of secondary antibody needed for the coupling procedure was approximately 43 µL. The washed MBs were suspended in 43 µL of PB with mixing. Then, 43 µL of goat anti-human IgM secondary antibody was added into the solution with thoroughly mixing. An equal volume, 43 µL of 1 M ammonium sulfate buffer was added into the solution with mixing and then the MB solution was incubated at 37 °C for 24 h. After the incubation, the MBs were separated from the solution using the magnet separation procedure. Then, the MBs were washed with 1 mL phosphate buffered saline  four times. The MBs were, finally, suspended in 330 µL of PBS with a concentration of 1 × 109 beads/mL. The secondary antibody coated MBs were stored at 4 to 8 °C with 0.02% (w/v) sodium azide  to provide a long shelf-life for the MCIA antibody test. The recommended ratio between the secondary antibodies and the MBs from the user manual was used in this study and the antibody coupling efficiency has been verified by coupling MBs to anti-human IgM secondary antibody-HRP instead of the goat anti-human IgM. This provided evidence that the secondary antibodies were successfully captured by the MBs using the recommended ratio. For the antibody test, the concentration of the secondary antibody conjugated MBs was optimized and should be higher than 5 × 106 beads/mL. The storage conjugated MBs were washed with PBS containing 0.1% bovine serum albumin (BSA)  once and then diluted by PBS to the desired concentration. A total of 10 µL of diluted conjugated MBs were resuspended in a 600 µL centrifuge vial. 10 µL of SARS-CoV-2 RBD IgM humanized coronavirus monoclonal antibodies  were added into the vial and mixed with a pipette tip for 30 s. After a 25 min incubation, during which SARS-CoV-2 IgM antibodies were captured by the IgM secondary antibodies, the conjugated MBs were sonicated for 10 s and then 10 μL of SARS-CoV-2 spike S1-His recombinant protein  with a concentration of 25 mg/L was added into the vial and mixed for 30 s. The aggregates were formed in this incubation procedure. After a 3 min incubation, the assay mixtures were introduced into the microflow cytometer by the external syringe pump. The flow rate for sample and sheath flows were set as 200 µL/h and 800 µL/h, respectively. A total of 2 min of data were collected and analyzed by the LabView program. For the SARS-CoV-2 IgG test, goat anti-rabbit IgG (H + L) antibodies  were coupled to the MBs instead. SARS-CoV-2 spike S1 antibodies  were tested by the MCIA. It is worth mentioning that the described immune reaction step of the MCIA can be easily automated, which is important for its POC applications.The MCIA is derived from the particle counting immunoassay with the principle that agglutination caused by an immune reaction decreases the number of free particles in assay mixtures . Microflx-axis is the time in milliseconds and the y-axis represents the amplitude of the signal. Red solid lines are waveforms of the signals. Peaks in x-axis in y-axis in Compared with monomers, aggregates have longer transit times. x-axis is the concentration of SARS-CoV-2 IgM and the y-axis is the average transit time. The stored goat anti-human IgM secondary antibody coated MBs were used to detect SARS-CoV-2 IgM on different days. Different coloured symbols, including black squares, red triangles and blue circles, are used in 2 was 0.9984, 0.9982 and 0.9941, respectively. The LOD of SARS-CoV-2 IgM for the MCIA was determined as 3.3 times the residual standard deviation of the regression line divided by the slope. Day 1 data were used to calculate the LOD, which was determined to be 0.06 mg/L. Iyer et al. reported a LOD of 0.28 mg/L for anti-RBD SARS-CoV-2 IgM for their in-house ELISA method, which is comparable to the LOD achieved by the MCIA [the MCIA . Additiothe MCIA . Therefox-axis is the concentration of SARS-CoV-2 IgG antibody and the y-axis is the average transit time. Goat anti-rabbit IgG (H + L) secondary antibody coated MBs with different storage times were used for the detection of SARS-CoV-2 IgG. Different coloured symbols including black squares, red triangles and blue circles, are used in 2 for each curve was consistent; for days 1, 8 and 15, the R2 was 0.9981, 0.9963 and 0.9987, respectively. The LOD of SARS-CoV-2 IgG for the MCIA was determined as 3.3 times the residual standard deviation of the regression line divided by the slope. Day 1 data were used to calculate the LOD, which was determined to be 0.10 mg/L. Iyer et al. reported a LOD of 0.04 mg/L for anti-RBD SARS-CoV-2 IgG for their in-house ELISA method, which is comparable to the LOD achieved by the MCIA [the MCIA . In addithe MCIA . TherefoAccording to the results presented in spiked sampleC is the concentration of the sample after the addition of a concentrated standard, which was diluted by the human serum. unspiked sampleC is the concentration of the sample prepared with PBS. addedC is the concentration of the added standard in the whole volume.To test the performance of the MCIA for its practical application, a certain number of SARS-CoV-2 antibodies diluted by human serum  were spiked into the samples and then tested by the MCIA. The recovery index was calculated by the following equation, where addedC was calculated to be 0.4 mg/L. unspiked sampleC was diluted by PBS to a concentration of 0.4 mg/L. In this case, spiked sampleC should be 0.8 mg/L, provided that the recovery index was 100%. After measurement of the samples, the concentrations of the samples were obtained from the calibration curve of the SARS-CoV-2 IgM day 1 measurement (Y = 24.964 × X + 105.73), which was shown in For the detection of SARS-CoV-2 IgM, 16 µL samples prepared by PBS were split into two equals. One was added with 2 µL of deionized water and the other was added with 2 µL of the concentrated standard of SARS-CoV-2 IgM. Both samples were measured by the MCIA. The concentrated standard was diluted by the human serum sample and The proposed MCIA can be used for the quantitative detection of SARS-CoV-2 antibodies and other kinds of antibodies as well. Compared with other POC antibody tests such as LFIAs, the MCIA can provide quantitative results instead of qualitative ones. The quantitative measurement of antibodies provides more information regarding the degree of the immune response, which may be helpful for monitoring the progression of COVID-19. The assay time of the MCIA is about half an hour, which meets the requirement of a POC test. The 10 µL sample volume consumption of the MCIA is comparable to most POC devices. Although the cost per analysis of the MCIA is dependent on the reagents needed, the MCIA is based on the microflow cytometry format, so the inherent advantages of microflow cytometry, such as cost-efficiency, also exist for the MCIA. The linear range of antibody concentrations detected by the MCIA is restricted by the prozone effect, but dilution can be performed when the concentration of the sample exceeds this range. Furthermore, it is important to note that there is about a 75% similarity between the genomes of SARS-CoV and SARS-CoV-2 [A MCIA has been developed for the POC quantitative detection of SARS-CoV-2 IgM and IgG antibodies. A linear relationship has been established between the average transit time and the concentration of SARS-CoV-2 antibodies up to 1.0 mg/L. 0.06 mg/L and 0.10 mg/L are the LODs for SARS-CoV-2 IgM and IgG, respectively. These values are comparable to those reported for in-house ELISAs, indicating that the device could adequately perform if tested against serum samples. The proposed protocol has demonstrated to be rapid, simple and sensitive for the POC quantitative measurement of SARS-CoV-2 antibodies. The 30 min assay time, 10 µL sample consumption, relatively simple procedures and compact core devices make the MCIA a promising tool for the POC detection of SARS-CoV-2 IgM and IgG. Furthermore, the developed protocol is readily adaptable to be applied for the detection of other antibodies. However, there are still some limitations of this study. The detection ranges of SARS-CoV-2 IgM and IgG were not large enough for the method to be conceivably performed without dilution of serum samples. The manual steps for the immune reaction need to be integrated into the MCIA to improve the detection efficiency. Further investigation of this study should focus on reducing the mentioned limitations and testing the sensitivity and specificity of the MCIA performance using serum specimens from individuals who have been infected with SARS-CoV-2.


Introduction: Aortic valve stenosis is the most frequent cardiac valve pathology in the western world. In high-risk patients, conventional aortic valve replacement (C-AVR) carries high rates of morbidity and mortality. In the last few years, rapid-deployment valves (RDV) have been developed to reduce the surgical risks. In this work, we aimed to compare the mid-term outcomes of rapid-deployment AVR (RD-AVR) with those of the C-AVR in high-risk patients.Methods: This retrospective case-control study identified 23 high-risk patients who underwent RD-AVR between 12/2015 to 01/2018. The study group was compared with a control group of 46 patients who were retrospectively selected from a database of 687 C-AVR patients from 2016 to 2017 which matched with the study group for age and Euro SCORE II.Results: RD-AVR group presented more cardiovascular risk factors. Euro SCORE II was higher in the RD-AVR group (P =0.06). In the RD-AVR group, we observed significantly higher mean prosthetic size (P <0.001). In-hospital mortality was zero in RD-AVR group versus 2 deaths in C-AVR group. Hospital stay was longer in the RD-AVR group with statistical significance (P =0.03). In the group AVR with associated cardiac procedures, while comparing subgroups RD-AVR versus C-AVR, early mean gradient was lower in the first cited (P =0.02). The overall mean follow-up was 10.9 ± 4.3 months.Conclusion: The RD-AVR technique is reliable and lead to positive outcomes. This procedure provides a much larger size with certainly better flow through the aortic root. It is an alternative to C-AVR in patients recognized to be surgically fragile. In some patients, the decision to use the RD-AVR was made peroperatively because of anatomical and pathological findings like huge valve calcification and small aortic root that were making conventional AVR (C-AVR) impossible or risky to perform. The study group was compared with a control group of 46 patients who were retrospectively selected from a database of 687 C-AVR patients from 2016 to 2017 which matched with the study group for age and EuroSCORE II . 10 : native aortic valve resection, sizing, and valve implantation by inverted and interrupted pledgeted suture with Cardioxyl® 2/0 . For the RD-AVR procedure, the aortic orifice was measured with the original sizer of the bioprosthesis. Three 4-0 polypropylene guiding sutures were passed at the nadir of the aortic annulus. The three guiding sutures were passed through the three holes arising from the annular ring of the prosthesis. Once the delivery system is in position, the valve is deployed by turning the release screw and placing the valve in place. Then the delivery system is removed. The field was rinsed with warm saline, and the prosthesis was dilated according to the recommendations.11 After closing of the aortotomy, transesophageal echocardiography was performed to assess the correct implantation of the prosthesis and the presence of any leakage.The C-AVR procedure is performed as fully described in our previous work http://cran.r-project.org). All tests were two-sided, with a Type I error set at 0.05. Patients were retrospectively matched  according to age, gender, EuroSCORE II and associated gesture. This was made by the method of the nearest neighbor. Categorical parameters were expressed as frequencies and associated percentages and continuous data as the mean ± standard deviation or as median [interquartile range], according to the statistical distribution. Categorical variables were compared between independent groups (RD-AVR or C-AVR) using the chi-square test or the Fisher exact test. Quantitative data were compared between groups with the Student t test or with the Mann-Whitney test, as appropriate. The Gaussian distribution was verified by the Shapiro-Wilk test and homoscedasticity by the Fisher-Snedecor test. Adjustments for age, associated gesture or EuroSCORE II were also considered in a multivariate point of view . Interactions between these factors and the two groups of patients (RD-AVR or C-AVR) were also studied. Subgroup analyses were then performed.Statistical analysis was performed using Stata software  and R 3.3.1 (P = 0.002). EuroSCORE II was higher in the RD-AVR group: 2.9  vs. 2.2  respectively, for a median of the RD-AVR and C-AVR (P = 0.06).P < 0.001). The most frequent associated procedure was CABG in both groups. There was no difference in term of CPB and cross-clamp time in both groups.P = 0.55). There was no death in the RD-AVR group. Both patients died in the ICU, one for multiorgan failure on the same day of operation and the other one suffered from sudden cardiac arrest on postoperative day 9 despite cardiopulmonary resuscitation and emergency resternotomy. P = 0.02, P = 0.03). On discharge echography, there was some mild/trivial periprosthetic leak (17.4%) in the RD-AVR group, but not requiring reoperation (P = 0.08). The mean gradient was slightly lower in the same group (P = 0.14). Regarding the subgroup AVR with associated cardiac procedures, the early postoperative mean gradient was 8.0 ± 3.2 mm Hg with the RD-AVR group and 11.3 ± 4.9 mm Hg C-AVR group (P = 0.02). All patients were free from aortic regurgitation > grade 2.In-hospital mortality rate was low (2.9%) and concerned 2 patients in the conventional group .Follow-up was completed in 100% of the patients. During the mean follow-up of 10.9 ± 4.3 months, there were no deaths, LVEF remained normal and mean gradient lower in the RD-AVR group . C-AVR in patients with a small aortic annulus may result in the prosthesis mismatch which is more common in smaller patients, obese and elderly women with multiple comorbidities.13 This procedure provides a much larger size with certainly better flow through the aortic root as the mean prosthetic size was greater in this group.4Although EuroSCORE II confirmed the higher score in the RD-AVR group compared to the control group, the postoperative course went well as no death was noticed in this risky group  and informed consent was taken from all the patients.None.